CN103467425A - Method for separating quercetin from oxytropis glabra - Google Patents
Method for separating quercetin from oxytropis glabra Download PDFInfo
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- CN103467425A CN103467425A CN2013104038677A CN201310403867A CN103467425A CN 103467425 A CN103467425 A CN 103467425A CN 2013104038677 A CN2013104038677 A CN 2013104038677A CN 201310403867 A CN201310403867 A CN 201310403867A CN 103467425 A CN103467425 A CN 103467425A
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- deionized water
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Abstract
The invention discloses a method for separating quercetin from oxytropis glabra, and the method comprises the following steps: (1) crushing oxytropis glabra; (2) extracting a total oxytropis glabra extract by using an ultrasonic assistance method; (3) separating oxytropis glabra flavones by using a macroporous resin adsorption method; (4) preparing a coarse quercetin product by virtue of acid hydrolysis; (5) recrystallizing quercetin; (6) detecting the purity of the quercetin. The method disclosed by the invention has the characteristics of simple preparation process, low cost and high product purity, and is suitable for promotion and application.
Description
Technical field
The invention belongs to medicine and health product technology field, relate to a kind of from glabrous crazyweed the method for separating meletin.
Background technology
Glabrous crazyweed (Oxytropis glabra DC) is pulse family (Leguminosae) Genus Oxytropis Dc (Oxytropis) plant, widely distributed in Xinjiang.According to anaesthetic allusion quotation record, glabrous crazyweed is a kind of important middle anaesthetic raw material, has anesthesia, calmness and analgesic effect, cures mainly toothache, arthrodynia, insomnia, forgetful, neurasthenia and skin pruritus.As glabrous crazyweed 4.5g, periploca sepium 6g and ground Chinese holly leaf 9g, be decocted in water for oral dose, can control arthrodynia.
Flavonoid compound is one of effective constituent of glabrous crazyweed, wherein contains Quercetin.Quercetin (quercetin), chemistry is by name 3,3 ', 4 ', 5,7-pentahydroxyflavone (3,3 ', 4 ', 5,7-pentahydroxyflavone), have another name called quercetin, Quercetin, can be used as medicine.Have preferably eliminate the phlegm, antitussive action, and certain antiasthmatic effect is arranged.Also have in addition anti-oxidant and remove free radical, protect cardiovascular, prevention coronary heart disease, control myocardial ischemia-reperfusion injury, anti-inflammatory, antianaphylaxis, the effects such as antiviral and calm, antitumor, preventing cancer.Can be used for treating chronic bronchitis, coronary heart disease and hyperpietic are also had to auxiliary therapeutic action.Up to the present, the patent application of separating meletin in glabrous crazyweed is not arranged.
Summary of the invention
The object of the invention is to overcome the defect that above-mentioned technology exists, a kind of method and application of separating meletin from glabrous crazyweed is provided, the method is beneficial to operation, product purity is high.
Its concrete technical scheme is:
A kind of from glabrous crazyweed the method for separating meletin, comprise the following steps:
(1) glabrous crazyweed is pulverized: get the glabrous crazyweed over-ground part, dry in the shade, pulverize, standby after mistake 20 mesh sieves;
(2) the Ultrasound-assisted method is extracted the glabrous crazyweed general extractive: take the glabrous crazyweed sample, the methyl alcohol ultrasonic extraction 90min that adds volume fraction 100% according to the ratio of material ratio 1:15g/mL, ultrasonic power 800W, temperature 45 C, filtration after extraction completes, filtrate adopts Rotary Evaporators concentrated, and the thermostatic drying chamber drying, obtain the glabrous crazyweed general extractive;
(3) separation of glabrous crazyweed flavones: glabrous crazyweed general extractive water is fully dissolved; To the deionized water that adds 1/3 volume in chromatography column, then D-101 type macroporous adsorbent resin is transferred in chromatography column with deionized water, add 70% ethanolic soln, volume is 2 times of resin volume, flow velocity is 1 times of resin volume/min, then with deionized water, is washed till without carrying out the gradient elution of the glabrous crazyweed general extractive aqueous solution after the alcohol flavor; Macroporous resin optimal adsorption processing condition are: pH value 4.0, temperature room temperature, flow velocity 1.0BV/h; Ethanol elution with 10%～100%, collect 50%～85% ethanol eluate, and concentrated through Rotary Evaporators, the freeze drier drying, obtain the brown color powder;
(4) separation of Quercetin crude product: take the about 1.0g of glabrous crazyweed flavones, adding volume ratio is V
methyl alcohol: V
water: V
hydrochloric acidthe hydrolyzed solution 60mL of=35:5:10, in 80 ℃ of heating in water bath 90min, cooled and filtered, 60 ℃ of oven dry obtain the thick product of yellow needle-like Quercetin;
(5) recrystallization of Quercetin: get the about 1.0g of Quercetin crude product prepared by step (4), add 100mL sherwood oil and 20mL deionized water, in 80 ℃ of heated and stirred 30min, filtered while hot, precipitate 3 times with the 20mL deionized water wash, in 60 ℃ of oven dry, obtain Quercetin; Take Quercetin (1) 0.5g, add the 100mL methanol solution, in 80 ℃ of dissolvings, filter, filtrate lets cool crystallization, filters to obtain crystal, with the 20mL deionized water wash, precipitates 3 times, in 60 ℃, dries to obtain the high Quercetin of purity.
Compared with prior art, beneficial effect of the present invention is:
The method of the invention is simple to operate, and organic solvent consumption is few, and production cost is low, process stabilizing, and energy consumption is little, pollutes less, and products obtained therefrom purity is high, applicable to laboratory and preparation of industrialization.Detection by Quercetin purity: the detection by ignition residue inspection, clarity detects and carries out purity detecting to separating the Quercetin obtained with high performance liquid chromatography, and result shows to separate the Quercetin purity obtained > 98%.
The accompanying drawing explanation
Fig. 1 is Quercetin HPLC figure.
Embodiment
Below in conjunction with the drawings and specific embodiments, technical scheme of the present invention is described in more detail
Glabrous crazyweed extracts test:
Influence factor for the extraction of integrated survey glabrous crazyweed flavones, utilize the Orthogonal Method characteristics of " the balanced dispersion ", choose ultrasonic time, methanol concentration, material ratio as the investigation factor, and choose again 3 level arrangement tests in conjunction with the every factor of experience of repeatedly extracting, the yield of flavones of take be to be weighed the objective indicator of extraction efficiency, preferably optimum extraction process.
Table 1 orthogonal experimental design
Table 2 glabrous crazyweed flavones extracts test-results
According to the optimum level that in orthogonal test, the K value of maximum is selected in each factor as a result, be: methanol concentration 100%, ultrasonic time 90min, solid-liquid ratio 1:15.
Under certain temperature, pH, the glabrous crazyweed extracting solution by the resin chromatography column, is measured the content of total flavones component in effluent liquid with certain flow velocity; Respectively sample solution flow velocity, temperature, pH are carried out to single factor experiment, determine the optimal adsorption processing condition.
The impact of table 3 sample solution pH value on macroporous resin adsorption
The impact of table 4 sample solution temperature on macroporous resin adsorption
The impact of table 5 sample solution flow velocity on macroporous resin adsorption
According to the single factor experiment result, and shorten as much as possible test period, definite optimal adsorption processing condition are: pH value 4.0, temperature room temperature, flow velocity 1.0BV/h.
The preparation of Quercetin needs comparatively fierce hydrolysising condition, yet condition is controlled the improper degraded that very easily causes again the flavones ring.Respectively composition, material ratio, hydrolysis time and four principal elements of temperature of hydrolyzed liquid are carried out respectively to one-factor experiment.
Table 6 hydrolyzed solution forms the impact on Quercetin purity
The impact of table 7 material ratio on Quercetin purity
The impact of table 8 hydrolysis temperature on Quercetin purity
The impact of table 9 hydrolysis time on Quercetin purity
According to single factor experiment result and experimentation cost, optimum test condition is: hydrolyzed solution forms V
methyl alcohol: V
water: V
hydrochloric acid=35:5:10, hydrolysis time 90min, solid-liquid ratio 1:60g/mL; 80 ℃ of hydrolysis temperatures.
Get Quercetin sample 1.5g and be placed in crucible, add sulfuric acid 1.0mL and dissolve, after low-temperature heat to sulfuric acid volatilizes, 550 ℃ of complete ashing, in the dislocation moisture eliminator, let cool to room temperature, accurately weighed after, obtain.Quercetin calcination slag is 0.038%, meets Ministry of Health's Medicine inspection standard (≤0.05%); The Quercetin sample is made into to 2% methanol solution, places and still clarify in one week, more do not melt the inspection of quality testing checking method by the tetrabutylammonium nonaqueous titrations, all clarify at ammonia solution, water and dehydrated alcohol, all meet the requirements; Quercetin HPLC chromatographic condition is: octadecylsilane bonding glue silica gel is weighting agent, and moving phase is acetonitrile-methyl alcohol (90:10), and flow velocity 1mL/min detects wavelength 226nm, 30 ℃ of column temperatures; Detection method: precision takes Quercetin 1.0mg, adds methyl alcohol 10mL that it is fully dissolved, then by methanol constant volume to 25mL, the HPLC sample size is 10 μ L, adopts normalization method working sample purity, as a result its purity > 98%.
The preparation method of Quercetin flavones in a kind of glabrous crazyweed, comprise the following steps: precision takes the glabrous crazyweed grass meal 1000.00g5 part crushed, according to optimum extraction process, extract, the method separating meletin of step (3)-(5), obtain respectively Quercetin crystallization 10.423mg, 10.532mg, 10.630mg, 10.445mg, 10.533mg; As shown in Figure 1, according to the HPLC method, detect, find that Quercetin purity is respectively 98.62%, 99.01%, 98.84%, 98.40%, 98.74%, average purity is 98.72%, and the variation coefficient is 2.63%.Result shows: this preparation method has accuracy preferably, extracts the Quercetin purity obtained higher.
The above; it is only preferably embodiment of the present invention; protection scope of the present invention is not limited to this; anyly be familiar with those skilled in the art in the technical scope that the present invention discloses, the simple change of the technical scheme that can obtain apparently or equivalence are replaced and are all fallen within the scope of protection of the present invention.
Claims (1)
1. the method for a separating meletin from glabrous crazyweed, is characterized in that, comprises the following steps:
(1) glabrous crazyweed is pulverized: get the glabrous crazyweed over-ground part, dry in the shade, pulverize, standby after mistake 20 mesh sieves;
(2) the Ultrasound-assisted method is extracted the glabrous crazyweed general extractive: take the glabrous crazyweed sample, the methyl alcohol ultrasonic extraction 90min that adds volume fraction 100% according to the ratio of material ratio 1:15g/mL, ultrasonic power 800W, temperature 45 C, filtration after extraction completes, filtrate adopts Rotary Evaporators concentrated, and the thermostatic drying chamber drying, obtain the glabrous crazyweed general extractive;
(3) separation of glabrous crazyweed flavones: glabrous crazyweed general extractive water is fully dissolved; To the deionized water that adds 1/3 volume in chromatography column, then D-101 type macroporous adsorbent resin is transferred in chromatography column with deionized water, add 70% ethanolic soln, volume is 2 times of resin volume, flow velocity is 1 times of resin volume/min, then with deionized water, is washed till without carrying out the gradient elution of the glabrous crazyweed general extractive aqueous solution after the alcohol flavor; Macroporous resin optimal adsorption processing condition are: pH value 4.0, temperature room temperature, flow velocity 1.0BV/h; Ethanol elution with 10%～100%, collect 50%～85% ethanol eluate, and concentrated through Rotary Evaporators, the freeze drier drying, obtain the brown color powder;
(4) separation of Quercetin crude product: take the about 1.0g of glabrous crazyweed flavones, adding volume ratio is V
methyl alcohol: V
water: V
hydrochloric acidthe hydrolyzed solution 60mL of=35:5:10, in 80 ℃ of heating in water bath 90min, cooled and filtered, 60 ℃ of oven dry obtain the thick product of yellow needle-like Quercetin;
(5) recrystallization of Quercetin: get the about 1.0g of Quercetin crude product prepared by step (4), add 100mL sherwood oil and 20mL deionized water, in 80 ℃ of heated and stirred 30min, filtered while hot, precipitate 3 times with the 20mL deionized water wash, in 60 ℃ of oven dry, obtain Quercetin; Take Quercetin (1) 0.5g, add the 100mL methanol solution, in 80 ℃ of dissolvings, filter, filtrate lets cool crystallization, filters to obtain crystal, with the 20mL deionized water wash, precipitates 3 times, in 60 ℃, dries to obtain the high Quercetin of purity.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104068211A (en) * | 2014-06-14 | 2014-10-01 | 塔里木大学 | Preparation method and application of oxytropis glabra DC flavone |
| CN104560399A (en) * | 2014-12-16 | 2015-04-29 | 湖北中烟工业有限责任公司 | Method for purifying jasmine flower extract for cigarettes |
| CN104774189A (en) * | 2015-03-31 | 2015-07-15 | 浙江大学 | Method for preparing high-purity quercetin |
| CN107746398A (en) * | 2017-10-23 | 2018-03-02 | 苏州鑫旷新材料科技有限公司 | A kind of method for spending middle extraction Quercetin from sunflower |
Citations (2)
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| WO1999062480A2 (en) * | 1998-05-29 | 1999-12-09 | Parfums Christian Dior | Use of at least a cosmetically acceptable saponin or sapogenol as cosmetic agent for increasing the amount of collagen iv in the dermal-epidermal junction |
| CN103202872A (en) * | 2013-04-08 | 2013-07-17 | 塔里木大学 | Preparation method and applications of flavonoid from Oxytropis glabra DC |
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2013
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999062480A2 (en) * | 1998-05-29 | 1999-12-09 | Parfums Christian Dior | Use of at least a cosmetically acceptable saponin or sapogenol as cosmetic agent for increasing the amount of collagen iv in the dermal-epidermal junction |
| CN103202872A (en) * | 2013-04-08 | 2013-07-17 | 塔里木大学 | Preparation method and applications of flavonoid from Oxytropis glabra DC |
Non-Patent Citations (2)
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| 于荣敏 等: "小花棘豆中黄酮醇和黄酮醇甙的分离鉴定", 《中国中药杂志》 * |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104068211A (en) * | 2014-06-14 | 2014-10-01 | 塔里木大学 | Preparation method and application of oxytropis glabra DC flavone |
| CN104560399A (en) * | 2014-12-16 | 2015-04-29 | 湖北中烟工业有限责任公司 | Method for purifying jasmine flower extract for cigarettes |
| CN104560399B (en) * | 2014-12-16 | 2017-11-07 | 湖北中烟工业有限责任公司 | A kind of purification process of cigarette jasmine flower extract |
| CN104774189A (en) * | 2015-03-31 | 2015-07-15 | 浙江大学 | Method for preparing high-purity quercetin |
| CN107746398A (en) * | 2017-10-23 | 2018-03-02 | 苏州鑫旷新材料科技有限公司 | A kind of method for spending middle extraction Quercetin from sunflower |
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